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The aim was to establish the chromatographic fingerprint of Rhizoma Panacis Japonici for its quality evaluation. Analysis was performed on a YMC-Pack ODS-AQ column (4.6 mm x 250 mm, 5 microm) eluted with the mobile phases of acetonitrile(A) and 5% acetic acid solution (B) at in gradient a flow rate of 1.0 mL x min(-1). The elution program was as follow: 0-5 min, changed from 20% A to 40% A, 5-20 min, kept 40% A. The temperature of column was 30 degrees C. The temperate of drift tube was 40 degrees C, and the nitrogen pressure was 33 Psi. Ten batches of Rhizoma Panacis japonici were determined. The HPLC-ELSD chromatographic fingerprint of chemical constituents was established from the 10 batches of Rhizoma Panacis japonici and it had 9 characteristic common peaks. The 10 batches of samples were classified into 2 cluster by cluster analysis. Furthermore, five known chemical constuituents were identified after isolation and purification by means of silica gel column chromatography, and semi-preparative high performance liquid chromatography. They were panaxsaponins Re, chikusetsusaponins IV, IVa, V and pjs-2, respectively. This study provided experimental data for comprehensive evaluation of the quality of Rhizoma Panacis japonici.